Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite

Juan David Ospina-Villa, Nancy Guillén, Cesar Lopez-Camarillo, Jacqueline Soto-Sanchez, Esther Ramirez-Moreno, Raul Garcia-Vazquez, Carlos A. Castañon-Sanchez, Abigail Betanzos, Laurence A. Marchat

Research output: Contribution to journalArticle

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Abstract

© 2017, The Microbiological Society of Korea and Springer-Verlag GmbH Germany. The 25 kDa subunit of the Clevage Factor Im (CFIm25) is an essential factor for messenger RNA polyadenylation in human cells. Therefore, here we investigated whether the homologous protein of Entamoeba histolytica, the protozoan responsible for human amoebiasis, might be considered as a biochemical target for parasite control. Trophozoites were cultured with bacterial double-stranded RNA molecules targeting the EhCFIm25 gene, and inhibition of mRNA and protein expression was confirmed by RT-PCR and Western blot assays, respectively. EhCFIm25 silencing was associated with a significant acceleration of cell proliferation and cell death. Moreover, trophozoites appeared as larger and multinucleated cells. These morphological changes were accompanied by a reduced mobility, and erythrophagocytosis was significantly diminished. Lastly, the knockdown of EhCFIm25 affected the poly(A) site selection in two reporter genes and revealed that EhCFIm25 stimulates the utilization of downstream poly(A) sites in E. histolytica mRNA. Overall, our data confirm that targeting the polyadenylation process represents an interesting strategy for controlling parasites, including E. histolytica. To our best knowledge, the present study is the first to have revealed the relevance of the cleavage factor CFIm25 as a biochemical target in parasites.
Original languageAmerican English
Pages (from-to)783-791
Number of pages703
JournalJournal of Microbiology
DOIs
StatePublished - 1 Oct 2017

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Entamoeba histolytica
Trophozoites
Polyadenylation
Parasites
Poly A
Messenger RNA
Bacterial RNA
Communicable Disease Control
Amebiasis
Double-Stranded RNA
Gene Targeting
Korea
Reporter Genes
Germany
Proteins
Cell Death
Western Blotting
Cell Proliferation
Polymerase Chain Reaction

Cite this

Ospina-Villa, Juan David ; Guillén, Nancy ; Lopez-Camarillo, Cesar ; Soto-Sanchez, Jacqueline ; Ramirez-Moreno, Esther ; Garcia-Vazquez, Raul ; Castañon-Sanchez, Carlos A. ; Betanzos, Abigail ; Marchat, Laurence A. / Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite. In: Journal of Microbiology. 2017 ; pp. 783-791.
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abstract = "{\circledC} 2017, The Microbiological Society of Korea and Springer-Verlag GmbH Germany. The 25 kDa subunit of the Clevage Factor Im (CFIm25) is an essential factor for messenger RNA polyadenylation in human cells. Therefore, here we investigated whether the homologous protein of Entamoeba histolytica, the protozoan responsible for human amoebiasis, might be considered as a biochemical target for parasite control. Trophozoites were cultured with bacterial double-stranded RNA molecules targeting the EhCFIm25 gene, and inhibition of mRNA and protein expression was confirmed by RT-PCR and Western blot assays, respectively. EhCFIm25 silencing was associated with a significant acceleration of cell proliferation and cell death. Moreover, trophozoites appeared as larger and multinucleated cells. These morphological changes were accompanied by a reduced mobility, and erythrophagocytosis was significantly diminished. Lastly, the knockdown of EhCFIm25 affected the poly(A) site selection in two reporter genes and revealed that EhCFIm25 stimulates the utilization of downstream poly(A) sites in E. histolytica mRNA. Overall, our data confirm that targeting the polyadenylation process represents an interesting strategy for controlling parasites, including E. histolytica. To our best knowledge, the present study is the first to have revealed the relevance of the cleavage factor CFIm25 as a biochemical target in parasites.",
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Ospina-Villa, JD, Guillén, N, Lopez-Camarillo, C, Soto-Sanchez, J, Ramirez-Moreno, E, Garcia-Vazquez, R, Castañon-Sanchez, CA, Betanzos, A & Marchat, LA 2017, 'Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite', Journal of Microbiology, pp. 783-791. https://doi.org/10.1007/s12275-017-7259-9

Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite. / Ospina-Villa, Juan David; Guillén, Nancy; Lopez-Camarillo, Cesar; Soto-Sanchez, Jacqueline; Ramirez-Moreno, Esther; Garcia-Vazquez, Raul; Castañon-Sanchez, Carlos A.; Betanzos, Abigail; Marchat, Laurence A.

In: Journal of Microbiology, 01.10.2017, p. 783-791.

Research output: Contribution to journalArticle

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T1 - Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite

AU - Ospina-Villa, Juan David

AU - Guillén, Nancy

AU - Lopez-Camarillo, Cesar

AU - Soto-Sanchez, Jacqueline

AU - Ramirez-Moreno, Esther

AU - Garcia-Vazquez, Raul

AU - Castañon-Sanchez, Carlos A.

AU - Betanzos, Abigail

AU - Marchat, Laurence A.

PY - 2017/10/1

Y1 - 2017/10/1

N2 - © 2017, The Microbiological Society of Korea and Springer-Verlag GmbH Germany. The 25 kDa subunit of the Clevage Factor Im (CFIm25) is an essential factor for messenger RNA polyadenylation in human cells. Therefore, here we investigated whether the homologous protein of Entamoeba histolytica, the protozoan responsible for human amoebiasis, might be considered as a biochemical target for parasite control. Trophozoites were cultured with bacterial double-stranded RNA molecules targeting the EhCFIm25 gene, and inhibition of mRNA and protein expression was confirmed by RT-PCR and Western blot assays, respectively. EhCFIm25 silencing was associated with a significant acceleration of cell proliferation and cell death. Moreover, trophozoites appeared as larger and multinucleated cells. These morphological changes were accompanied by a reduced mobility, and erythrophagocytosis was significantly diminished. Lastly, the knockdown of EhCFIm25 affected the poly(A) site selection in two reporter genes and revealed that EhCFIm25 stimulates the utilization of downstream poly(A) sites in E. histolytica mRNA. Overall, our data confirm that targeting the polyadenylation process represents an interesting strategy for controlling parasites, including E. histolytica. To our best knowledge, the present study is the first to have revealed the relevance of the cleavage factor CFIm25 as a biochemical target in parasites.

AB - © 2017, The Microbiological Society of Korea and Springer-Verlag GmbH Germany. The 25 kDa subunit of the Clevage Factor Im (CFIm25) is an essential factor for messenger RNA polyadenylation in human cells. Therefore, here we investigated whether the homologous protein of Entamoeba histolytica, the protozoan responsible for human amoebiasis, might be considered as a biochemical target for parasite control. Trophozoites were cultured with bacterial double-stranded RNA molecules targeting the EhCFIm25 gene, and inhibition of mRNA and protein expression was confirmed by RT-PCR and Western blot assays, respectively. EhCFIm25 silencing was associated with a significant acceleration of cell proliferation and cell death. Moreover, trophozoites appeared as larger and multinucleated cells. These morphological changes were accompanied by a reduced mobility, and erythrophagocytosis was significantly diminished. Lastly, the knockdown of EhCFIm25 affected the poly(A) site selection in two reporter genes and revealed that EhCFIm25 stimulates the utilization of downstream poly(A) sites in E. histolytica mRNA. Overall, our data confirm that targeting the polyadenylation process represents an interesting strategy for controlling parasites, including E. histolytica. To our best knowledge, the present study is the first to have revealed the relevance of the cleavage factor CFIm25 as a biochemical target in parasites.

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